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检测肺组织以及支气管肺泡灌洗细胞和灌洗液中的脂质过氧化反应。

Detection of lipid peroxidation in lung and in bronchoalveolar lavage cells and fluid.

作者信息

Petruska J M, Wong S H, Sunderman F W, Mossman B T

机构信息

Department of Pathology, University of Vermont College of Medicine, Burlington 05405-0068.

出版信息

Free Radic Biol Med. 1990;9(1):51-8. doi: 10.1016/0891-5849(90)90049-o.

Abstract

Inhalation of toxic materials such as asbestos, silica, 100% oxygen, ozone, or nitrogen dioxide may lead to an increased production of reactive oxygen metabolites which may initiate lipid peroxidation. Measurement of lipid peroxidation in cells and fluid obtained by bronchoalveolar lavage (BAL), as well as in lung tissue, may aid in monitoring the development and extent of pulmonary damage after inhalation of a toxic substance. In this study, we employed a sensitive assay for detection of malondialdehyde (MDA), a breakdown product of lipid peroxidation. By separation of the adduct with thiobarbituric acid, using a reverse phase high pressure liquid chromatographic technique, we accurately and sensitively measured the content of MDA in BAL cells, lavage fluid, and lavaged lung tissue homogenates of rats. The amounts of sample required for detection of MDA were small enough possibly to be applied to use with human specimens; in addition, recovery of added MDA was acceptable with all types of samples. Inclusion of a metal chelator in the preparation of samples appeared necessary to prevent metal-catalyzed propagation of lipid peroxidation during the assay. Overall, the method described here using samples from rats may be applicable to detecting lipid peroxidation in BAL samples from humans.

摘要

吸入石棉、二氧化硅、纯氧、臭氧或二氧化氮等有毒物质可能会导致活性氧代谢产物生成增加,进而引发脂质过氧化。通过支气管肺泡灌洗(BAL)获取的细胞和液体以及肺组织中脂质过氧化的测量,可能有助于监测吸入有毒物质后肺损伤的发展和程度。在本研究中,我们采用了一种灵敏的检测方法来检测脂质过氧化的分解产物丙二醛(MDA)。通过使用反相高压液相色谱技术将加合物与硫代巴比妥酸分离,我们准确且灵敏地测量了大鼠BAL细胞、灌洗液和灌洗肺组织匀浆中MDA的含量。检测MDA所需的样品量足够小,有可能应用于人体标本;此外,所有类型的样品中添加的MDA回收率都可以接受。在样品制备中加入金属螯合剂似乎有必要,以防止在检测过程中金属催化脂质过氧化的传播。总体而言,这里描述的使用大鼠样品的方法可能适用于检测人类BAL样品中的脂质过氧化。

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